JPS60130601A - Manufacture of hyaluronic acid water-insoluble composition - Google Patents
Manufacture of hyaluronic acid water-insoluble compositionInfo
- Publication number
- JPS60130601A JPS60130601A JP59162779A JP16277984A JPS60130601A JP S60130601 A JPS60130601 A JP S60130601A JP 59162779 A JP59162779 A JP 59162779A JP 16277984 A JP16277984 A JP 16277984A JP S60130601 A JPS60130601 A JP S60130601A
- Authority
- JP
- Japan
- Prior art keywords
- hyaluronic acid
- carried out
- crosslinking agent
- treatment
- water
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
- C08B37/0072—Hyaluronic acid, i.e. HA or hyaluronan; Derivatives thereof, e.g. crosslinked hyaluronic acid (hylan) or hyaluronates
Abstract
(57)【要約】本公報は電子出願前の出願データであるた
め要約のデータは記録されません。(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.
Description
【発明の詳細な説明】
この発明はヒアルロン酸又はその塩の生体適合性で水圧
不溶な組成物及びその製法に関する。こＱ発明の組成物
は、生体適合性であるので、人工心臓パル′プや、導管
移植片などを含む人工器官のような各種の生体内用途忙
用いることができる。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a biocompatible and hydraulically insoluble composition of hyaluronic acid or a salt thereof and a method for producing the same. Because the compositions of the present invention are biocompatible, they can be used in a variety of in-vivo applications, such as artificial heart pulps, prostheses, including ductal grafts, and the like.
また、この発明の組成物は、各種の重合体製品の改質に
Ｊ伯いることができ、この改質品は同様に各種生体内用
途を有する。Furthermore, the composition of the present invention can be used to modify various polymer products, and the modified products likewise have various in vivo applications.
ヒアルロン酸は、公知の天然物であって、医薬や生物学
で多くの用途が知られているもので（たとえば米国特許
第４．２ ’７２，５２２号及びその中の引用文献参照
）、またヒアルロン酸のある種の架橋ゲμは公知−ｃ”
６る（ａ−１ｚンら、ＡＱｔａ Ｃｈｅｍ、 ５ｃａｎ
ａ。Hyaluronic acid is a well-known natural product with many uses in medicine and biology (see, for example, U.S. Pat. No. 4.2'72,522 and references cited therein); Certain cross-linking molecules of hyaluronic acid μ are known.
6ru (a-1zn et al., AQta Chem, 5can
a.
１８ １９６４ 陥ユ、ｐ２１４〜５）。18, 1964, Fallout, p.214-5).
この発明によれば、ヒアルロン酸又はその塩をホルムア
ルデヒド、ジメチロール尿素、ジメチロールエチレン尿
累、エチレンオキシド、ポリアジリジン、ポリインシア
ネート及びジビニルスルボンからなる群より選択された
架橋剤との処理に付すことからなる水不溶性ヒアルロン
酸又はその塩の組成物の製法が提供される。According to the present invention, hyaluronic acid or a salt thereof is treated with a crosslinking agent selected from the group consisting of formaldehyde, dimethylolurea, dimethylolethylene urea, ethylene oxide, polyaziridine, polyincyanate, and divinylsulfone. A method for producing a composition of water-insoluble hyaluronic acid or a salt thereof is provided.
この発明におけるヒアルロン酸又はその塩杜、市販品又
はそれから誘導されたものが用いることができる。ヒア
ルロン酸の塩は、生体適合性で多るかぎ）、各種の公知
の塩が含まれる。好ましい具体例としてナトリウム、カ
リシム、カｐシクムなどの塩が含まれる。In this invention, hyaluronic acid or its salts, commercially available products, or products derived therefrom can be used. Salts of hyaluronic acid include various known salts that are biocompatible. Preferred examples include salts such as sodium, calisim, capsicum, and the like.
ヒアルロン酸又はその塩は、架橋時に粉末状、フィルム
状、ゲル状の何れであってもよい。さらに、支持材で支
持されたものであってもよい。支持材としては、たとえ
ばポリエチレンテレフタレートのよ、うな合成樹脂の編
物のシートや、イオン交換樹脂、シリカ、アルミナのよ
うな粒状物であってもよい。粉状物を例にとれば、粉状
物を予めヒアルロン酸又はその塩で被覆し、得られる被
覆粉状物をこの発明の架橋剤で架橋されてもよい。Hyaluronic acid or a salt thereof may be in any form of powder, film, or gel during crosslinking. Furthermore, it may be supported by a support material. The support material may be a knitted sheet of synthetic resin, such as polyethylene terephthalate, or a particulate material such as ion exchange resin, silica, or alumina. Taking a powder as an example, the powder may be coated with hyaluronic acid or its salt in advance, and the resulting coated powder may be crosslinked with the crosslinking agent of the present invention.
また、編物のシートを文に材とする場合には、ヒアルロ
ン酸又はその塩はフィルム状として支持されたものであ
る。Furthermore, when a knitted sheet is used as the material, hyaluronic acid or its salt is supported in the form of a film.
この発明で使用する架橋剤は、前述のごとき特定Ｏもの
であるが、架橋条件は架橋剤の種類によって変えられる
。処理は乾燥又は水性条什下、室温又はＭ流温度で行う
ことができる。その１例を挙げると架橋剤がジビニルス
ルボンの場合、架橋約２０℃で行うことができる。ヒア
ルロン酸又はその塩は、熱に脱感でｂるのでなるべく低
いｍ度の処理が好ましい。The crosslinking agent used in this invention is a specific type as mentioned above, but the crosslinking conditions can be changed depending on the type of crosslinking agent. The treatment can be carried out under dry or aqueous conditions, at room temperature or at stream temperature. For example, when the crosslinking agent is divinyl sulfone, the crosslinking can be carried out at about 20°C. Since hyaluronic acid or its salt is desensitized by heat, it is preferable to treat it at a temperature as low as possible.
より詳しい処理条件は、次に挙げる実施例によって説明
する。More detailed processing conditions will be explained with reference to the following examples.
次にこの発明の数種のＡ体側を実施例（特に明記しない
限ハ重址部）で例証するが、これによってこの発明は限
定されるものではない。Next, several types of the A-body side of this invention will be illustrated by Examples (with heavy parts unless otherwise specified), but this invention is not limited thereby.
実施例１
水−アセトン混合物に、５ｔ９６Ｃ重量）ホルムアルデ
ヒド水溶液と濃塩酸を加えた。得られた混合物ｔｖ組成
（重ｆｉｋｇ６ ） ハ、ＣＨ，Ｏ０，２７； ＨＧＪ
０．１９；水／アセトン比ｌ：２８であった。ヒアル
ロン酸ナトリウム粉末（０，５９）を、この混合物５０
ｄ中で１０分間加熱還流した。次いで粉末をＦ取し、十
分に水／ア七トン１：３混液で洗い、次いでアセトンで
洗い、真空オープン中で乾燥した。生成したヒアルロン
酸粉末は、水に不溶で、結合したＣＨ２Ｏ′ｆｒ１．４
１９６含有した◎実施例２
次（Ｄｍ成（１ｉ９６ ） ： ＣＨ２Ｏｇ、５ ；Ｈ
Ｑｅ ｏ、：ｓａ ；水／アセトン比ｌ：２の架橋混液
で、上記の実施例Ｉｔ−緑如返し行った。生成物のＣ迅
０含量は５．３９６であった。Example 1 To a water-acetone mixture, an aqueous formaldehyde solution (5t96C weight) and concentrated hydrochloric acid were added. Obtained mixture tv composition (heavy fikg6) Ha, CH, O0,27; HGJ
0.19; water/acetone ratio l:28. Sodium hyaluronate powder (0,59) was added to this mixture for 50
The mixture was heated to reflux for 10 minutes in d. The powder was then filtered, thoroughly washed with a 1:3 mixture of water and acetone, then acetone and dried in an open vacuum. The produced hyaluronic acid powder is insoluble in water and has a combined CH2O'fr1.4
◎Example 2 containing 196 (Dm composition (1i96): CH2Og, 5; H
Qe o, :sa; The above Example It-green was carried out with a crosslinking mixture having a water/acetone ratio of 1:2. The C content of the product was 5.396.
実施例１と２で、ヒアルロン酸粉末の架橋は、水−アセ
トン漏液中で実施された。水／アセトン比とホルムアル
デヒド濃度を変化さすことによシ、生成物のｓａ比（ｓ
ｗｅｌｌｉｎｇ ｒａｔｉｏ ）をｉｎすることが可能
である。かくして、膨問比は実施例１の生成物で１７８
９６で実施例２のものは２３０９６であった。膨潤化は
混液中のアセトン証とホルムアルデヒド濃度とを増加さ
すことにより減少さすことができる。In Examples 1 and 2, crosslinking of hyaluronic acid powder was carried out in a water-acetone leak. By varying the water/acetone ratio and formaldehyde concentration, the sa ratio (s
welling ratio). Thus, the swelling ratio is 178 for the product of Example 1.
96, and that of Example 2 was 23,096. Swelling can be reduced by increasing the acetone and formaldehyde concentrations in the mixture.
次の実施例は、架橋剤としてポリアジリジン化合物を使
用する場合を例証する。このポリアジリジンタイプの化
合物は、乾燥状態で室温でとアルロン酸を架橋するが、
慮温で行うことはヒアルロン酸が熱に脱感なポリマーで
あるためヒアルロン酸の場合に特に重要である。The following example illustrates the use of polyaziridine compounds as crosslinking agents. This polyaziridine type compound crosslinks aruronic acid at room temperature in the dry state, but
Carrying out the treatment at a moderate temperature is particularly important in the case of hyaluronic acid, since hyaluronic acid is a heat-desensitizing polymer.
実施例３
と７７ｙａｙ酸の水溶液１１３．ｏｒ（５度１４．：ａ
ｒｒ、ｙ／―）に、 Ｏ，４５Ｖのポリアジリジン化合
物−クロス−リンカ−０Ｘ１ｏｏ （ポリビニルケミカ
ル社製）を加えた。架橋剤のヒアルロン酸に対するモル
比は０．５であった。混合物をガラス板で５＆ｉ＋厚さ
の層として、室温で２日間乾燥した。架信ヒアルロン酸
の透明フィルムが得られ、このものは水に溶解せず、か
つ１６０９６の水中の膨潤比を有した。Example 3 Aqueous solution of 77yay acid with 113. or(5 degrees 14.:a
rr, y/-), a polyaziridine compound-cross-linker-0X1oo (manufactured by Polyvinyl Chemical Co., Ltd.) of O,45V was added. The molar ratio of crosslinking agent to hyaluronic acid was 0.5. The mixture was dried in a 5&i+ thick layer on a glass plate for 2 days at room temperature. A transparent film of cross-linked hyaluronic acid was obtained, which was not soluble in water and had a swelling ratio in water of 16096.
実施例４
Ｏ１５１の乾燥ヒアルロン酸粉末を、クロヌーリンカ−
０Ｘ−１ｏｏの１９６アセトン液５０ ｔｔｔｌと混合
し、５分間保った後、戸取した。粉末を空気中で２時間
乾燥し、次いで水で数回洗い、真空オープン中４０℃で
４時間乾燥した。架橋粉末の水中での膨潤比は１３５９
（ｉであった。Example 4 Dry hyaluronic acid powder of O151 was added to Kuronu Linker.
It was mixed with 50 tttl of 196 acetone solution of 0X-1oo, kept for 5 minutes, and then taken out. The powder was dried in air for 2 hours, then washed several times with water and dried at 40° C. in an open vacuum for 4 hours. The swelling ratio of crosslinked powder in water is 1359
(It was i.
次の実施例は、高度の膨潤性を有する架橋ヒアルロン酸
を得るためボリアシリノン化合物を用いた場合を例証す
るものである。The following example illustrates the use of boriasilinone compounds to obtain crosslinked hyaluronic acid with a high degree of swelling.
実施例５
固形のとアＡ／μン酸ナトリクム０．６ｔを、ｏ、ｅ９
６クロスーリンカーＣＸ−１００の水溶液と混合した。Example 5 0.6 t of solid ToA/μ sodium chloride was added to o, e9
6 was mixed with an aqueous solution of cross-linker CX-100.
得うレ六−Ｉ８液はヒアルロン酸す−トリウムに列する
０Ｘ−１００のモＡ／比＝０．１を有した。ｒＵ液中ノ
ヒアμロン酸ナトリウム含蓋は６．０４０６ （ＭＲ）
であった。得られた高粘稠性混合液のｌ）Ｈを２％塩酸
で２，５に調製した。得られたフィルムは水に易溶性で
あった。このフィルムを６０℃で３０分間加熱した。こ
の加熱処理で強固な水不溶性のフィルムが得らハた。The resulting liquid 6-I8 had an 0.times.100 moA/ratio of 0.1, which is on the order of thorium hyaluronate. Sodium uronate content in rU solution is 6.0406 (MR)
Met. The l)H of the resulting highly viscous mixture was adjusted to 2.5 with 2% hydrochloric acid. The obtained film was easily soluble in water. This film was heated at 60°C for 30 minutes. This heat treatment resulted in a strong water-insoluble film.
実施例６
繊維様のヒアルロン酸ナトリウム（０，１０９：Ｍ）を
１９６ポリイノシアナート（モーベイ・ケミカル社製、
デスモジュアＮ−７５）のアセトン１７１ Ｋ ２５ｕ
ｌと混合し、混合物を１０分間還流した。沈澱物を分離
し、アセトンで３回洗浄し、４５ｍＨ／の減圧下６０℃
で３０分間乾燥し、最後に５酸化燐入り減圧デシケータ
−で乾燥した。得られた生成１７（ａｌ１２７ｔ） は
水に不溶で、１２０９（ｉの膨潤比を有した。Example 6 Fiber-like sodium hyaluronate (0,109:M) was mixed with 196 polyinocyanate (manufactured by Mauvay Chemical Co., Ltd.,
Desmodua N-75) acetone 171 K 25u
The mixture was refluxed for 10 minutes. The precipitate was separated, washed three times with acetone, and heated at 60 °C under a vacuum of 45 mH/
for 30 minutes, and finally dried in a vacuum desiccator containing phosphorus pentoxide. The resulting product 17(al127t) was insoluble in water and had a swelling ratio of 1209(i).
実施例７
とアルロン酸ナトリクム水溶液（濃度９．８ Ｗ／ｄ
）の６ｔを、 Ｏ，０１７ｒのＮ、Ｎ−ジメチロールエ
チレン尿素及び０．００５９の硝酸亜鉛と混合した。混
合物をガラス板上にギャヌトし、−夜装置乾燥した。Example 7 and sodium aluronate aqueous solution (concentration 9.8 W/d
) was mixed with O,017r of N,N-dimethylolethylene urea and 0.0059 of zinc nitrate. The mixture was poured onto a glass plate and dried in an apparatus overnight.
得られたフィルムを１１０℃で１５分間加熱処理した。The obtained film was heat-treated at 110° C. for 15 minutes.
そのものは、水に不溶で、１４５９（５の膨潤比を示し
た。It was insoluble in water and exhibited a swelling ratio of 1459 (5).
次の実施例はヒアルロンｒａを架橋するためにノビニル
スルホンを用いた場合である。The following example uses novinyl sulfone to crosslink hyaluronic acid ra.
実施例８
０．４ｏ１ｔ （１ミリモ／Ｌ／）のジビニルスルホン
含有のヒアルロン酸ナトリウム溶液３５ｍｇにジピニｙ
ｙ ｙｖホンｏ、１ｘａｆ （ｌ ミ！Ｊ −６Ｎ ）
ｔ−７Ｊｌ；合した。Example 8 Dipiny was added to 35 mg of a sodium hyaluronate solution containing 0.4 o1t (1 mm/L/) of divinyl sulfone.
y yvhon o, 1xaf (l mi!J -6N)
t-7Jl; combined.
混合物のｐＨを１９６水酸化ナトリウム水溶液で約８．
５〜９に調整した。混合物をガラス板にキャストシ、−
夜室温で乾燥して、フィルムを得た。このフィルムは水
に易溶であった。フィルムを６０℃で３０分間加熱して
、強固な水に不溶なフィルムを得た。The pH of the mixture was adjusted to approximately 8.0 with 196 aqueous sodium hydroxide solution.
Adjusted to 5-9. Cast the mixture on a glass plate, −
A film was obtained by drying at room temperature overnight. This film was easily soluble in water. The film was heated at 60° C. for 30 minutes to obtain a strong water-insoluble film.
実箆例９
ＩＰ！４ｆｊヒアｐロン酸の乾燥フィルムを、２６２の
アセトンと１１の水との混液に０．６ｙのジビニＡ／ス
／Ｉ／ホンを溶解した液中に入れ、１０分間保持した。Real example 9 IP! A dry film of 4fj hyalpronic acid was placed in a solution of 0.6y diviny A/S/I/hon in a mixture of 262 parts acetone and 11 parts water and held for 10 minutes.
溶液からフィルムを取り出し、空気中で１０分間乾燥し
、次いでオーブン中６５℃で２０分間加熱した。ヒアル
ロン酸の強固な架橋フィルムが得られた。The film was removed from the solution, dried in air for 10 minutes, and then heated in an oven at 65° C. for 20 minutes. A strong crosslinked film of hyaluronic acid was obtained.
実施例１０
風乾ヒアルロン酸ナトリウム２．９５５’を０．２規定
水酸化すＩｌｌｌｌラム液５７．３５１と混合し、完全
に溶解するまでガラス棒で攪拌した。次いでジビニルヌ
ルホン１１をその混合物に混合攪拌し、室温で１時間放
置した。混合物は、堅いゲルとなった。このゲルを１０
０ｄの水とともに、Ｖｉｒ ＴｉＳゝ１４５“ホモゲナ
イザに入れ、３０．００Ｏｒｐｍで５分間処理した。非
常に膨潤した小さな粒子が得られた。この粒子を水で数
回洗浄し、ガフスロート上吸引ｐ取した。膨潤率を測定
するため、約１２のゲルを、１５１！Ｉｌのガラスフィ
ルターに入れ、次−でゲラステックの遠心管に入れ、３
．００Ｏｒｐｍで５０分間遠心分離した。圧出した水が
遠心管の底に集められた。ゲル中のヒアルロン酸濃度は
、０．２１％て、即ち水中での膨潤比が４７６でちった
。Example 10 2.955' of air-dried sodium hyaluronate was mixed with 57.351 of 0.2N hydroxide Illll Lamb's solution and stirred with a glass rod until completely dissolved. Next, divinylnurphone 11 was mixed and stirred into the mixture, and the mixture was allowed to stand at room temperature for 1 hour. The mixture became a stiff gel. 10 times this gel
It was placed in a Vir TiS 145'' homogenizer with 0 d of water and treated at 30.00 rpm for 5 min. Very swollen small particles were obtained. The particles were washed several times with water and vacuumed up the gaff throat. To measure the swelling ratio, approximately 12 gels were placed in a 151!Il glass filter, then placed in a Gelastec centrifuge tube at 3.
．． Centrifugation was performed at 000 rpm for 50 minutes. The extruded water was collected at the bottom of the centrifuge tube. The hyaluronic acid concentration in the gel was 0.21%, that is, the swelling ratio in water was 476.
実施例１１
実施例１０に記載した手法でゲ／ｌ／を得、これを０．
１５モル食塩水に分数じた。粒子を同じ食塩水で洗浄し
た。遠心分ｙｉ後のゲル中のヒアルロン酸濃度は１．２
９９６で、膨潤比は７７．５であった。Example 11 Ge/l/ was obtained by the method described in Example 10, and this was converted to 0.
The mixture was aliquoted into 15 molar saline. The particles were washed with the same saline solution. The hyaluronic acid concentration in the gel after centrifugation yi is 1.2
996, the swelling ratio was 77.5.
実Ｊム例１２
風乾ヒアルロン酸ナトリウム１．０９ ｆ ０．２モ／
’）Ｊ＜酸化ナトリクム水溶液９．Ｏｆｆに浴解し、ガ
フス棒で攪拌した。得られた粘稠液に、ジビニルスルホ
ン０．３３ｙを混合攪拌し、室温で２０時間放置しｆｃ
。Actual Example 12 Air-dried sodium hyaluronate 1.09 f 0.2 mo/
') J<Sodium oxide aqueous solution 9. The solution was dissolved in the bath off and stirred with a gaff rod. To the obtained viscous liquid, 0.33y of divinyl sulfone was mixed and stirred, and the mixture was left at room temperature for 20 hours.
.
得られた硬いゲ／Ｉ／を実施例１０と同様に処理しｌ〔
。The obtained hard gel /I/ was treated in the same manner as in Example 10.
.
遠心万緑後のゲル中のヒアルロン酸濃度は４．３０％で
あシ、即ち膨潤比が２３．６であった。The hyaluronic acid concentration in the gel after centrifugation was 4.30%, that is, the swelling ratio was 23.6.
この発明による製剤の生体適合性は、下記のゲスト法に
よシ例証される。The biocompatibility of the formulations according to the invention is illustrated by the Guest method below.
実五例１３−血液適合性テスト
実施例１０によって作られたサンプルの血液反応性を評
価する予備０［究に、与■−セロ）ニンのヒト血小板に
よる放出を用いた。正常ヒト静脈血をグツスナック・シ
リンジで採取し、直ちに３．８９６クエン酸ナトリウム
含有のグツスナック・チューブに移した（９部の全血に
対し１部のクエン酸ナトリウム）、。EXAMPLE 13 - BLOOD COMPATIBILITY TEST In a preliminary study to evaluate the blood reactivity of the samples made according to Example 10, the release of seronin by human platelets was used. Normal human venous blood was collected with a Gutsnak syringe and immediately transferred to a Gutsnak tube containing 3.896 sodium citrate (9 parts whole blood to 1 part sodium citrate).
血小板に富んだ血漿を、４℃で１２５ Ｘｇで１５分間
遠心分離で作り、血清学用ピペットでプラスチック製又
はシリコン化試験管に移した。血小板ｔｉｃ Ｎ ／ｖ
タ血Ｎｉｌ （ＰＥＰ ）、６．２〜０．５ ｔｔＥ
／ｍ ＰＦＪ’に、３Ｈ−セロトニン（”Ｈ−５−ヒト
ｌ’ＪＩｒＶ）リプタミン（”Ｈ−５ＨＴ）ニュー・イ
ングフンド・ヌクｖｙｆＭ、２６．３０ｉ／ｍｏｌ、
ｌ ｍ Ｃ１／ｄ！ 工ｊｒノーｐ−水〕を加え、３７
℃で１５分間培養した。この検定では、シリコン化又は
ボリグロビレン試験管ヲ用い、トロンビンを正のコント
ローμとして使用し、被覆したサングル及び被覆しない
サングルをテストした。検定すべきサン１μ含有の複数
の管の各々に、１．０〜２．Ｏｗｌの一−５Ｅ（Ｔ −
Ｐぴを加えた。全放射能を測定するためコントロール混
合物から５０μｅの部分標品を採取した。適当な培養期
間（１０〜１２０分）を経た後、懸濁液の０．２〜０．
５をとシ、エベンドμフ・小型遠心分離機中のシリコン
油上で１２，０ＯＯＸ（）で１０分間遠心分離する。容
管から５Ｑｄの上澄液をとり、５ｄＯ液体ンンテレーシ
ョン液を加え、放射量をベーター分光測定で測定した。Platelet-rich plasma was generated by centrifugation at 125 Xg for 15 minutes at 4°C and transferred to plastic or siliconized tubes with a serology pipette. Platelet tic N/v
Tag blood Nil (PEP), 6.2-0.5 ttE
/m PFJ', 3H-serotonin ("H-5-human l'JIrV) liptamine ("H-5HT) New Ingfund NukuvyfM, 26.30i/mol,
l m C1/d! Add water], 37
The cells were incubated at ℃ for 15 minutes. In this assay, coated and uncoated samples were tested using siliconized or polyglopylene test tubes, using thrombin as a positive control μ. 1.0-2. Owl One-5E (T-
Added Ppi. A 50 μe aliquot was taken from the control mixture to measure total radioactivity. After a suitable incubation period (10-120 minutes), the suspension is 0.2-0.
5 and centrifuged for 10 minutes at 12,000 OOX () over silicone oil in an Everend microcentrifuge. The supernatant liquid of 5Qd was taken from the container, 5dO liquid anteration liquid was added, and the amount of radiation was measured by beta spectroscopy.
トロンビン又はテストサンプルで放出された３Ｈ−５ｆ
（Ｔの量は、上澄液の放射量の増加（実験サンプルの放
射量−コントロールの放射量）である。テスト物質は、
１２０分まで”Ｈ−５ＨＴの顕著な血小板放出を誘因し
な力為った。3H-5f released in thrombin or test sample
(The amount of T is the increase in the radioactivity of the supernatant (the radioactivity of the experimental sample - the radioactivity of the control). The test substance is
Up to 120 minutes, H-5HT did not induce significant platelet release.
トイ 代理人 弁理士 野河信太邸toy Agent: Patent Attorney Shinta Nogawa House
Claims (1)
ジメチロール尿素、ジメチロールエチレン尿素、エチレ
ンオキシド、ポリアジリジン、ｄ！リイソシ”１ネート
およびジビニルスルホンからなる群から選択された架橋
剤との処理に付すことからなる水不溶性のヒアルロン酸
又はその塩の組成物の製法。 ２、架橋剤がホルムアルデヒドで、処理が水性媒体中還
流温度で行われる特許請求の範囲１項による方法。 ３、架橋剤がポリアジリジンで、処理が乾燥状態で定温
で行われる特許請求の範囲１項による方法。 ４、 １ＪｉＡ橋剤が４ζリイソＶアナートで１．処理
がアセトン中還流温度で行われる特許請求の範囲１項に
よる方法〇 ５、架橋剤がジメチロールエチレン尿素で、処理が約１
１０℃で行われる特許請求の範囲１項による方法。 ６、架橋剤がジビニルスルホンで、処理が約６０〜６５
℃で行われる特許請求の範囲１項による方法。 ７、架橋剤がジビニルスルホンで、処理が約２０℃でア
ルカリ条件下で行われる特許請求の範囲１項による方法
。 ８、 ヒアルロン酸又はその塩が、粉末状、フィル、ζ
状又はゲル状の形で架橋されてなる特許請求の範囲１〜
７項の何れかの方法。[Claims] 1. Hyaluronic acid or its salt, formaldehyde,
Dimethylol urea, dimethylol ethylene urea, ethylene oxide, polyaziridine, d! A method for producing a composition of water-insoluble hyaluronic acid or its salt, which comprises subjecting it to treatment with a crosslinking agent selected from the group consisting of lysocyanate and divinyl sulfone. 2. The crosslinking agent is formaldehyde and the treatment is carried out in an aqueous medium. 3. A method according to claim 1, which is carried out at a medium reflux temperature. 3. A method according to claim 1, in which the crosslinking agent is polyaziridine and the treatment is carried out in a dry state at a constant temperature. 4. The 1JiA crosslinking agent is 4ζlyiso 1. Process according to claim 1, wherein the treatment is carried out in acetone at reflux temperature, the crosslinking agent is dimethylolethylene urea, and the treatment is carried out in acetone at reflux temperature.
A process according to claim 1 carried out at 10°C. 6. The crosslinking agent is divinyl sulfone, and the treatment is about 60-65
A process according to claim 1 carried out at <0>C. 7. A process according to claim 1, wherein the crosslinking agent is divinyl sulfone and the treatment is carried out at about 20° C. under alkaline conditions. 8. Hyaluronic acid or its salt in powder form, fill, ζ
Claims 1~
Any method in Section 7.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US56181883A | 1983-12-15 | 1983-12-15 | |
| US561818 | 1983-12-15 | ||
| US598071 | 1984-04-09 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS60130601A true JPS60130601A (en) | 1985-07-12 |
Family
ID=24243605
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP59162779A Pending JPS60130601A (en) | 1983-12-15 | 1984-07-31 | Manufacture of hyaluronic acid water-insoluble composition |
Country Status (2)
| Country | Link |
|---|---|
| JP (1) | JPS60130601A (en) |
| DE (1) | DE3434082A1 (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS61164558A (en) * | 1985-01-17 | 1986-07-25 | 生化学工業株式会社 | Molding material of medical molded article |
| JPS61234864A (en) * | 1985-03-01 | 1986-10-20 | バイオマトリクス・インコーポレイテツド | Molded article and its production |
| JPS62129226A (en) * | 1985-11-29 | 1987-06-11 | バイオマトリツクス、インコ−ポレ−テツド | Drug release system depending on hyaluronane, derivatives ortheir salts and manufacture |
| JPH0669490B2 (en) * | 1984-07-23 | 1994-09-07 | カビ・フアーマシア・アー・ベー | Implant for preventing adhesion between body tissues and method for producing the same |
| JPH0669481B2 (en) * | 1984-06-08 | 1994-09-07 | カビ・フアーマシア・アー・ベー | Substitute vitreous humor consisting of cross-linked hyaluronic acid gel |
| WO1995031223A1 (en) * | 1994-05-13 | 1995-11-23 | Kuraray Co., Ltd. | Medical polymer gel |
| WO2007015579A1 (en) | 2005-08-04 | 2007-02-08 | Teijin Limited | Cellulose derivative |
| WO2009041627A1 (en) | 2007-09-28 | 2009-04-02 | Shiseido Company Ltd. | Swellable crosslinked hyaluronic acid powder and method for producing the same |
| US8137685B2 (en) | 2003-10-29 | 2012-03-20 | Teijin Limited | Hyaluronic acid compound, hydrogel thereof and joint treating material |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5202431A (en) * | 1985-07-08 | 1993-04-13 | Fidia, S.P.A. | Partial esters of hyaluronic acid |
| US4851521A (en) * | 1985-07-08 | 1989-07-25 | Fidia, S.P.A. | Esters of hyaluronic acid |
| IT1198449B (en) * | 1986-10-13 | 1988-12-21 | F I D I Farmaceutici Italiani | ESTERS OF POLYVALENT ALCOHOLS OF HYALURONIC ACID |
| WO2003089476A1 (en) * | 2002-04-17 | 2003-10-30 | Genzyme Corporation | Cross-linked hyaluronate compounds |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4152170A (en) * | 1975-06-18 | 1979-05-01 | Sumitomo Chemical Company, Ltd. | Cross-linked pullulan |
| GB1515963A (en) * | 1975-07-15 | 1978-06-28 | Massachusetts Inst Technology | Crosslinked collagen-mucopolysaccharide composite materials |
-
1984
- 1984-07-31 JP JP59162779A patent/JPS60130601A/en active Pending
- 1984-09-17 DE DE19843434082 patent/DE3434082A1/en active Granted
Non-Patent Citations (1)
| Title |
|---|
| ACTA CHEM.SCAND=1964 * |
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0669481B2 (en) * | 1984-06-08 | 1994-09-07 | カビ・フアーマシア・アー・ベー | Substitute vitreous humor consisting of cross-linked hyaluronic acid gel |
| JPH0669490B2 (en) * | 1984-07-23 | 1994-09-07 | カビ・フアーマシア・アー・ベー | Implant for preventing adhesion between body tissues and method for producing the same |
| JPS61164558A (en) * | 1985-01-17 | 1986-07-25 | 生化学工業株式会社 | Molding material of medical molded article |
| JPS61234864A (en) * | 1985-03-01 | 1986-10-20 | バイオマトリクス・インコーポレイテツド | Molded article and its production |
| JPS62129226A (en) * | 1985-11-29 | 1987-06-11 | バイオマトリツクス、インコ−ポレ−テツド | Drug release system depending on hyaluronane, derivatives ortheir salts and manufacture |
| US5770229A (en) * | 1994-05-13 | 1998-06-23 | Kuraray Co., Ltd. | Medical polymer gel |
| WO1995031223A1 (en) * | 1994-05-13 | 1995-11-23 | Kuraray Co., Ltd. | Medical polymer gel |
| US8137685B2 (en) | 2003-10-29 | 2012-03-20 | Teijin Limited | Hyaluronic acid compound, hydrogel thereof and joint treating material |
| WO2007015579A1 (en) | 2005-08-04 | 2007-02-08 | Teijin Limited | Cellulose derivative |
| US8378091B2 (en) | 2005-08-04 | 2013-02-19 | Teijin Limited | Cellulose derivative |
| WO2009041627A1 (en) | 2007-09-28 | 2009-04-02 | Shiseido Company Ltd. | Swellable crosslinked hyaluronic acid powder and method for producing the same |
| JP4460617B2 (en) * | 2007-09-28 | 2010-05-12 | 株式会社資生堂 | Swellable crosslinked hyaluronic acid powder and method for producing the same |
| KR100990301B1 (en) | 2007-09-28 | 2010-10-26 | 가부시키가이샤 시세이도 | Swellable crosslinked hyaluronic acid powder and its manufacturing method |
| JPWO2009041627A1 (en) * | 2007-09-28 | 2011-01-27 | 株式会社資生堂 | Swellable crosslinked hyaluronic acid powder and method for producing the same |
Also Published As
| Publication number | Publication date |
|---|---|
| DE3434082A1 (en) | 1985-07-11 |
| DE3434082C2 (en) | 1991-02-21 |
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